| 货号 | 12191S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/ChIP |
| 目标/特异性 | SPT16 (D7I2K) Rabbit mAb recognizes endogenous levels of total SPT16 protein. |
| 使用方法 | WB(1:1000) ChIP (1:50) |
| 供应商 | CST |
| 背景 | Supressor of Ty-16 (SPT16) and structure-specific recognition protein-1 (SSRP1) are subunits of the facilitates chromatin transcription (FACT) complex that is essential for transcription elongation (1,2). FACT facilitates RNA polymerase-dependent transcription of chromatin templates by destabilizing the nucleosomes within the open reading frames of active genes (3-5). FACT destabilizes the nucleosomes, which would otherwise act as barriers to RNA polymerase transcription activity, by disrupting histone-histone and histone-DNA contacts that lead to the eviction of the histone H2A-H2B dimer (2,3,6). FACT may also function as a histone chaperone to reassemble nucleosomes after RNA polymerase passage (7). In addition to transcription, FACT activity has been shown to have a role in DNA replication in yeast and in DNA repair by contributing to the activation of p53 by CK2 and by facilitating histone H2AX-H2B exchange upon DNA damage (8-10). |
| 存放说明 | -20C |
| 计算分子量 | 140 |
Western blot analysis of extracts from various cell lines using SPT16 (D7I2K) Rabbit mAb.Western blot 方法检测不同细胞系的提取物，使用的抗体为SPT16 (D7I2K) Rabbit mAb。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT 116 cells starved for 48 hr then serum stimulated with 10% FBS for 15 min and either 10 μl of SPT16 (D7I2K) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human c-Fos Promoter Primers #4663, SimpleChIP® Human c-Fos Exon 3 Primers #12010, SimpleChIP® Human EGR1 Promoter Primers #5549, and SimpleChIP® Human EGR1 Intron 3 Primers #11953. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.使用从4 x 106 HCT 116细胞中提取的交联过的染色质，以及10 µl SPT16 (D7I2K) Rabbit mAb 或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀，这些细胞需要饥饿培养48小时然后用10% FBS进行15分钟血清刺激。所用试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。富集到的DNA经过实时PCR定量，所使用引物为SimpleChIP® Human c-Fos Promoter Primers #4663, SimpleChIP® Human c-Fos Exon 3 Primers #12010, SimpleChIP® Human EGR1 Promoter Primers #5549, 以及SimpleChIP® Human EGR1 Intron 3 Primers #11953。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。 |
